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Non-coding RNA regulation of sex differences in stroke

非编码RNA对中风性别差异的调节

基本信息

批准号：
10401426
负责人：
Creed Michael Stary
金额：
$ 36.12万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10401426
关键词：
3&apos Untranslated Regions Acute Advanced Development Affect Animals Apoptosis Inhibitor Astrocytes Biological Biological Phenomena Brain Brain Injuries Cause of Death Cell Culture Techniques Cell Death Cell Survival Cell physiology Cells Cessation of life Clinical Clinical Treatment Clinical Trials Data Development Disease Elderly Evolution Experimental Models Failure Female Fluorescent in Situ Hybridization Functional disorder Gene Expression Regulation Gene Targeting Generations Genes Glucose Immunohistochemistry In Vitro Incidence Injury Intravenous Investigation Ischemia Knowledge Life Link Mediating MicroRNAs Middle Cerebral Artery Occlusion Mitochondria Molecular Mus Nerve Growth Factors Neurodegenerative Disorders Neurons Outcome Oxidative Phosphorylation Pathogenesis Pathway interactions Pharmaceutical Preparations Pharmacological Treatment Play Prevalence Process Proteins RNA immunoprecipitation sequencing RNA methylation Reactive Oxygen Species Recovery Regulation Regulator Genes Reperfusion Therapy Resolution Rodent Role SIRT1 gene Sex Differences Site Small Interfering RNA Source Stroke Testing Therapeutic Untranslated RNA Woman Work age related aged brain cell cell type cohort comparative design disability fluorescence imaging in vivo in vivo evaluation ischemic injury male men mitochondrial dysfunction novel post stroke pre-clinical pre-clinical research preclinical study reelin protein response response to injury sex sexual dimorphism stroke outcome stroke therapy translational barrier trend young adult

项目摘要

Stroke remains the second leading cause of death worldwide and the primary cause of long-term disability in the US. Although pre-clinical studies have identified hundreds of potential drug agents, the only pharmacological treatment available remains early reperfusion with thrombolytics. A major factor for translational failure may be the overwhelming use of young male rodents in pre-clinical research, and lack of sex-specific design and analysis in clinical trials. Stroke is a sexually dimorphic disease with sex differences in incidence, prevalence, and outcome. Mitochondria mediate post-ischemia cell death in both males and females, but in different manners. Non-coding RNAs, including microRNAs (miRs), are upstream regulators of genes that regulate cell survival and mitochondrial function. In young adult male animals, miRs have been established as central regulators in the cellular response to stroke, however their role in females or aged animal cohorts represents a critical knowledge gap. Our prior studies (2, 4) and preliminary evidence suggest age-related differences between females and males in expression of select miRs and their targets following stroke. The overarching hypothesis of these Aims is that expression and function of miR-181a and miR-200c is a critical determinant of sexual dimorphism in stroke outcomes. To test this, we will first compare aged (20 month old) female and male mice using the middle cerebral artery occlusion (MCAO) model of experimental stroke, then compare primary female and male neuronal and astrocyte cultures utilizing in vitro ischemia. Preliminary evidence in aged animals reveals only a modest post- MCAO miR-181a response that is limited to females, while comparatively, a pronounced miR-200c response occurs in both sexes. Moreover, our preliminary evidence implies sex-differences in miR-181a and miR-200c gene targeting. In Aim 1, we will first compare protection with post-MCAO IV treatment of anti-miR-181a or anti- miR-200c in aged males and females. Then, we will assess the long-term cell-specific and sex-specific response in miR-181a and miR-200c to MCAO. Next, we will compare sustained post-MCAO miR-200c inhibition on long- term neuro-recovery between aged males and females, with the basis that preliminary evidence implies female- specific targeting of the neurotrophic protein reelin by miR-200c. In Aim 2, we will: 1) focus on the roles of miR- 181a and miR-200c in differences in regional (core versus penumbra) post-stroke disruption in mitochondrial function between aged males and females; and, 2) determine the roles of miR-181a and miR-200c in cell-type, and sex-specific alterations in oxidative phosphorylation and disruptions in cytosolic and mitochondrial Ca2+- handling following in vitro ischemia. In Aim 3 we will first define cell-type and sex-dependent differential gene targeting by miR-181a (Grp78 and XIAP) and miR-200c (XIAP, sirtuin-1, reelin). Finally we will determine whether observed differential gene targeting could be attributed to differences in RNA methylation, and whether this changes in response to injury. Delineating the molecular mechanisms that determine sexual dimorphism in stroke is necessary to overcome translational barriers and advance the development of novel stroke treatments.

中风仍然是世界范围内第二大死亡原因，也是老年人长期残疾的主要原因。我们尽管临床前研究已经确定了数百种潜在的药物制剂，但唯一的药理学可用的治疗仍然是用溶血栓剂进行早期再灌注。翻译失败的主要因素可能是临床前研究中大量使用年轻雄性啮齿动物，并且缺乏性别特异性设计和分析在临床试验阶段脑卒中是一种两性异形疾病，在发病率、患病率和死亡率方面存在性别差异。结果。线粒体介导缺血后细胞死亡在男性和女性，但在不同的方式。非编码RNA，包括微小RNA（miR），是调节细胞存活和细胞增殖的基因的上游调节因子。线粒体功能在年轻的成年雄性动物中，miR已被确定为细胞中的中央调节因子。细胞对中风的反应，然而，它们在雌性或老年动物队列中的作用代表了关键的知识间隙我们之前的研究（2，4）和初步证据表明，女性和男性之间的年龄相关差异，男性在中风后选择miR及其靶点的表达。这些目标的首要假设 miR-181 a和miR-200 c表达和功能是中风性二型性的关键决定因素结果。为了测试这一点，我们将首先使用大脑中区比较老年（20个月大）雌性和雄性小鼠，动脉闭塞（MCAO）模型的实验性中风，然后比较初级女性和男性的神经元，利用体外缺血的星形胶质细胞培养物。在老年动物中的初步证据显示， MCAO miR-181 a反应仅限于女性，而相对而言，miR-200 c反应明显两性都有。此外，我们的初步证据表明miR-181 a和miR-200 c存在性别差异，基因靶向在目标1中，我们将首先比较MCAO后IV治疗的抗miR-181 a或抗miR-181 a的保护作用。老年男性和女性中的miR-200 c。然后，我们将评估长期的细胞特异性和性别特异性反应 miR-181 a和miR-200 c与MCAO的关系。接下来，我们将比较MCAO后miR-200 c的持续抑制对长时间的老年男性和女性之间的长期神经恢复，初步证据表明女性- miR-200 c特异性靶向神经营养蛋白reelin。在目标2中，我们将：1）关注miR-1的作用， 181 a和miR-200 c在脑卒中后线粒体区域（核心与半暗带）破坏中的差异 2）确定miR-181 a和miR-200 c在细胞类型中的作用，氧化磷酸化的性别特异性改变以及细胞质和线粒体Ca 2 +- 体外缺血后的处理。目的3首先定义细胞类型和性别依赖的差异基因通过miR-181 a（Grp 78和XIAP）和miR-200 c（XIAP，sirtuin-1，reelin）靶向。最后，我们将确定观察到的差异基因靶向可以归因于RNA甲基化的差异，以及这是否对伤害的反应。描述了决定两性异形的分子机制，中风是克服转化障碍和促进新中风治疗发展所必需的。